Medium data

Search for medium no. = [1030].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1030   DISSULFURIMICROBIUM HYDROTHERMALE MEDIUM

NH4Cl  0.33    g
KCl  0.33    g
MgCl2·6H2O  0.33    g
CaCl2·2H2O  0.33    g
KH2PO4  0.33    g
Trace elements solution (see below)  10.0     ml
Sulfur (powder)  5.0     g
Distilled water  800.0     ml
Mix components, except sulfur, bring to a boil and cool down under a CO2 (100%) gas stream. While gassing, add the following components to the medium:
Trace vitamin (see Medium No. 197)  10.0     ml
NaHCO3  2.0     g
Bubble CO2 for 3 - 4 min and check pH to be 6.5 - 6.8. Readjust pH to the range, if necessary. Then, suspend ferrihydrite (see below) in the medium and dispense the suspension into culture vessels (e.g., 10 ml in Hungate tubes) under a CO2 gas stream. Add appropriate amount of sulfur (powder). Seal the culture vessels with butyl rubber stoppers and autoclave at 105C for 1 hr.
Ferrihydrite: Slowly titrate 320 ml of FeCl3·6H2O solution (60.0 g/L) with 10% (w/v) NaOH to pH 7.0-7.5 with agitation on a magnetic stirrer (approximately 80-100 ml NaOH solution). Remove ferrihydrite into a container, cover with water and store at room temperature overnight. Remove supernatant after centrifugation at 2000 rpm for 5 min.
Trace elements solution:
(NH4)2Fe(SO4)2·6H2O  78.4     mg
CoCl2·6H2O  23.8     mg
NiCl2·6H2O  16.6     mg
Na2MoO4·2H2O  2.42    mg
Na2WO4·2H2O  5.49    mg
ZnSO4·7H2O  14.4     mg
CuCl2·2H2O  0.17    mg
Na2SeO4  9.45    mg
H3BO3  0.62    mg
MnCl2·4H2O  19.8     mg
SrSO4  0.92    mg
CrK(SO4)2·12H2O  0.5     mg
AlCl3·6H2O  2.41    mg
BaCl2  2.08    mg
Na2SiO3·9H2O  14.2     mg
KBr  11.9     mg
KI  16.6     mg
Na2SO4  71.0     mg
Distilled water  1.0     L